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One of the main clinical manifestations presented by victims of snake bite envenoming are coagulation disorders. Considering that fibrinogen is a key molecule for crosslinked fibrin clot formation, the objective of this work was the quantitative analysis of the fibrinogenolytic activity of snakes of medical importance in Brazil and neutralization by specific antivenom. For this, pools of three genera of medical importance (Bothrops, Crotalus and Lachesis) that are used for the production of antivenom were used, and three pools of species of the genus Bothrops that are not part of the pool for the production of antivenom. The Lachesis pool had the highest fibrinogenolytic activity, even demonstrating partial cleavage (42.9 % consumption) of the fibrinogen gamma chain. The Bothrops genus venom pools have shown subtle variations between them. The Crotalus pool, despite not showing total cleavage of any fibrinogen chain, began cleavage of fibrinogen by the beta chain. The specific antivenoms used were able to delay the cleavage of fibrinogen in all the venoms used, which could be the first step towards implementing previous in vitro tests to analyze the quality of the batches of antivenoms produced, thus potentially reducing the use of animals used in this process., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests or personal relationships that could appear to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Naja kaouthia is a medically important snake, widely distributed throughout Southeast Asia, with a diverse venom composition. N. kaouthia venom is subject to significant intraspecific variation, caused by several factors, such as the wide geographic distribution of the species, sexual and ontogenetic factors. However, individual variation is a factor that has only been studied with small sample size groups and/or with pooled samples. With this in mind, this study evaluates the composition and in-vitro enzymatic activities of 29 individual venom samples from specimens born in captivity, with a similar genetic background caused by inbreeding, using SDS-PAGE under reducing conditions, RP-HPLC profiles and enzymatic activities of PLA 2 , LAAO and proteolytic activity over azocasein. Even in this scenario, we were able to observe significant variations in abundance and activity of PLA 2 . Individual variations in LAAO activity, as well as a sexual dimorphism in which males present a significantly higher LAAO activity than females were observed. Phosphodiasterase and CRiSP abundance were also found and considered to have multiple effects in the clinical manifestations of envenomation by presenting synergistic effects with other proteins from N. kaouthia venom. The RP-HPLC profiles were better at detecting compositional differences than SDS-PAGE profiles and better correlated with enzymatic activities, being a better technique to screen variation profiles and reinforcing the importance of individual venom analysis prior to pooling., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Venom-induced hemorrhage analysis usually is performed by Minimum Hemorrhagic Dose (MHD), however a similar method can be used to compare venoms with fewer laboratory animals. Our work compared the MHD of five different venoms, with the size of hemorrhagic spot, finding good correlations in the results. Considering the 3Rs principle, we propose the use of the hemorrhagic spot method to compare hemorrhagic activity of snake venoms, rather than using the MHD method, since the first one needs 5 times less animals than the other., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Fasciolosis is a parasitosis of great importance for livestock, as well as for public health, as it is considered by the WHO as a neglected disease. Disease control is complex and reinfections make the use of therapeutic products an unsustainable method from an economic, environmental and health point of view. The aim of this study was to evaluate a new vaccine formulation for dairy cattle, containing soluble Fasciola hepatica antigens associated with Montanide 763 AVG and saponin adjuvants (FhSAMS). The vaccine was tested with two protocols, a single dose and a booster dose 6 months after the first dose. The FhSAMS vaccine proved to be safe, with no side effects. Furthermore, it was able to generate a more robust humoral immune response when a six-month booster dose was used, in addition to stimulating greater production of IFN-ʏ, indicating a Th1 profile immune stimulus., (© 2024 John Wiley & Sons Ltd.)

Background: Although My Jump 2 ® consistently presented excellent values of reliability and validity when compared to force platforms (FPs) and contact mats, to date no scientific investigation assessed the validity and reliability of My Jump 2 ® to measure jump height in visually impaired five-a-side soccer athletes. Thus, the study aimed at analyzing the validity and reliability of the My Jump 2 ® to measure the vertical jump of five-a-side soccer athletes., Methods: Twelve visually impaired five-a-side soccer athletes, volunteered for this study. Each player performed five countermovement jumps (CMJs) and squat jumps (SJs) on a contact platform (CP) while they were simultaneously recorded using My Jump 2 ® ., Results: There was almost perfect agreement between the My Jump 2 ® and the contact platform measurements of CMJ (intraclass correlation coefficient = 0.99; p < 0.001) and SJ (intraclass correlation coefficient = 0.99; p < 0.001), heights for athletes during the first and second measurement days. Bland-Altman analysis showed a bias of 0.25 ± 0.5 cm; maximum SD = 1.3; minimum SD = -0.88 for CMJ, while that Bland-Altman analysis showed bias 0.18 ± 0.5 cm; maximum SD = 1.3; minimum SD = -0.96, for SJ., Conclusion: We can conclude that the My Jump 2 ® is a valid and reliable method to measure CMJ and SJ in visually impaired five-a-side soccer athletes., Competing Interests: Alexandre Igor Araripe Medeiros is an Academic Editor for Peerj., (© 2024 Silva et al.)

The genus Bothrops are considered Category 1 of medical importance by the World Health Organization, responsible for approximately 85 % of snakebites occurring throughout Brazil. Main factors determining snake venom variations can be genetics, diet, gender, geographic distribution, age, or even seasonality. In this study, we compared the composition of protein profile, biochemical activities, and immunorecognition of toxins present in the venom of eight adults of Bothrops species ( B. alternatus, B. atrox, B. jararaca, B. jararacussu, B. leucurus, B. moojeni, B. neuwiedi and B. pauloensis ). The following methods were used to analyze the venoms: protein dosage; electrophoresis in polyacrylamide gel containing SDS; High Performance Liquid Chromatography - Reverse Phase; enzymatic activities, western blotting and Enzyme Linked Immuno Sorbent Assay. The results show inter and intraspecific differences in the electrophoretic profile. LAAO and PLA 2 activities, in general, were higher in males than females and proteolytic activity was higher in females than males. The bothropic antivenom produced by Instituto Butantan recognized most of the protein bands in all Bothrops species analyzed, with only the regions between 37 and 25 kDa presenting lower intensity. A notable variability in the chromatograms was observed. Bothrops venom demonstrated inter-intraspecific disparities in protein composition and biochemical activity., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Thais Almeida de Godoy reports financial support was provided by Sao Paulo State Department for Health Coordination of Science Technology and Strategic Health Supplies. Anita Mitico Tanaka-Azevedo reports was provided by State of Sao Paulo Research Foundation. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Rattlesnakes belonging to the genus Crotalus are widely distributed throughout the Americas. In Brazil, symptoms commonly associated with envenomation by Crotalus durissus collilineatus include myalgia, rhabdomyolysis, renal failure, neurotoxicity, and progressive paralysis, which are related to the protein composition of this venom. Snake venom composition exhibits compositional variability that may reflect geographic distribution, age, captivity, diet, sex, and even individual genetics. Although seasonality is also considered a possible source of variation, there are few reports of such variability in snake venom. In this work, venoms of the same eight C. durissus collilineatus were extracted every three months for two years, to analyze seasonal changes in composition and activities. To this end, venom composition was analyzed by protein quantification, SDS-PAGE, and HPLC, and the LAAO, PLA 2 and coagulant activities were measured. Venoms of these C. d. collilineatus showed minor seasonal differences in venom activities and no composition differences were found. LAAO and coagulant activities displayed a pattern of seasonal change, while PLA 2 activity seemed to have no seasonality tendency. Also, there are sexual differences, in which males seem to be more stable than females in regard to some activities. Individual variability occurs even in seasonal variation of activities, highlighting the importance of controlling circumstances of venom extraction before comparing results between groups of snakes., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Anita Mitico Tanaka Azevedo reports financial support was provided by State of Sao Paulo Research Foundation. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Antarctica harbors a microbial diversity still poorly explored and of inestimable biotechnological value. Cold-adapted microorganisms can produce a diverse range of metabolites stable at low temperatures, making these compounds industrially interesting for biotechnological use. The present work investigated the biotechnological potential for antimicrobial and antitumor activity of filamentous fungi and bacteria isolated from marine sediment samples collected at Deception Island, Antarctica. A total of 89 microbial isolates were recovered from marine sediments and submitted to an initial screening for L-glutaminase with antitumoral activity and for antimicrobial metabolites. The isolates Pseudogymnoascus sp. FDG01, Pseudogymnoascus sp. FDG02, and Penicillium sp. FAD33 showed potential antiproliferative action against human pancreatic carcinoma cells while showing no toxic effect on non-tumor cells. The microbial extracts from unidentified three bacteria and four filamentous fungi showed antibacterial activity against at least one tested pathogenic bacterial strain. The isolate FDG01 inhibited four bacterial species, while the isolate FDG01 was active against Micrococcus luteus in the minimal inhibitory concentration of 0.015625 μg mL  -1 . The results pave the way for further optimization of enzyme production and characterization of enzymes and metabolites found and reaffirm Antarctic marine environments as a wealthy source of compounds potentially applicable in the healthcare and pharmaceutical industry., (© 2024. The Author(s), under exclusive licence to Springer Nature Japan KK, part of Springer Nature.)

Microorganisms that inhabit the cold Antarctic environment can produce ligninolytic enzymes potentially useful in bioremediation. Our study focused on characterizing Antarctic bacteria and fungi from marine sediment samples of King George and Deception Islands, maritime Antarctica, potentially affected by hydrocarbon influence, able to produce enzymes for use in bioremediation processes in environments impacted with petroleum derivatives. A total of 168 microorganism isolates were obtained: 56 from sediments of King George Island and 112 from Deception Island. Among them, five bacterial isolates were tolerant to cell growth in the presence of diesel oil and gasoline and seven fungal were able to discolor RBBR dye. In addition, 16 isolates (15 bacterial and one fungal) displayed enzymatic emulsifying activities. Two isolates were characterized taxonomically by showing better biotechnological results. Psychrobacter sp. BAD17 and Cladosporium sp. FAR18 showed pyrene tolerance (cell growth of 0.03 g mL -1 and 0.2 g mL -1 ) and laccase enzymatic activity (0.006 UL -1 and 0.10 UL -1 ), respectively. Our results indicate that bacteria and fungi living in sediments under potential effect of hydrocarbon pollution may represent a promising alternative to bioremediate cold environments contaminated with polluting compounds derived from petroleum such as polycyclic aromatic hydrocarbons and dyes., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

In Brazil, the genus Bothrops is responsible for most ophidian accidents. Snake venoms have a wide variety of proteins and peptides exhibiting a broad repertoire of pharmacological and toxic effects that elicit systemic injury and characteristic local effects. The snakes' natural resistance to envenomation caused by the presence of inhibitory compounds on their plasma have been extensively studied. However, the presence of these inhibitors in different developmental stages is yet to be further discussed. The aim of this study was to evaluate the ontogeny of Bothrops jararaca plasma inhibitor composition and, to this end, plasma samples of B. jararaca were obtained from different developmental stages (neonates, youngs, and adults) and sexes (female and male). SDS-PAGE, Western blotting, affinity chromatography, and mass spectrometry were performed to analyze the protein profile and interaction between B. jararaca plasma and venom proteins. In addition, the presence of γBjPLI, a PLA2 inhibitor previously identified and characterized in B. jararaca serum, was confirmed by Western blotting. According to our results, 9-17% of plasma proteins were capable of binding to venom proteins in the three developmental stages. The presence of different endogenous inhibitors and, more specifically, different PLA2 inhibitor (PLI) classes and antihemorrhagic factors were confirmed in specimens of B. jararaca from newborn by mass spectrometry. For the first time, the αPLI and βPLI were detected in B. jararaca plasma, although low or no ontogenetic and sexual correlation were found. The γPLI were more abundant in adult female, than in neonate and young female, but similar to neonate, young and adult male according to the results of mass spectrometry analysis. Our results suggest that there are proteins in the plasma of these animals that can help counteract the effects of self-envenomation from birth., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Serino-Silva et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

This work presents the characterization of a novel naturally phosphorylated starch extracted from an unconventional and non-utilized source, the seeds of the stone fruit Syzygium malaccense. The morphology and chemical characteristics of the extracted starch were examined by scanning electron microscopy, FTIR, 1 H/ 13 C/ 31 P NMR and 13 C-CP/MAS-NMR, HPAEC-PAD chromatography, XRD, DSC, and RVA. The extraction yielded a highly pure starch (95.6 %) with an average granule size of 13 μm. The analysis of the starch components revealed an amylose content of 28.1 % and a predominance (65 %) of B-chains (B1-B3 65 %) in the amylopectin, as shown through HPAEC-PAD chromatography. The X-ray diffractogram was compatible with B-type starch, which was confirmed by the deconvolution of the C1 peak in the 13 C-CP/MAS-NMR. X-Ray diffractogram also showed that S. malaccense has 28.5 % of crystallinity. DSC analysis showed values of 82.6 °C and -12.41 J g -1 for Tc and ΔH, respectively, which is compatible with a highly ordered starch granule structure. The values observed for peak (4678 mPa•s), trough (3055 mPa•s), and final viscosity (6526 mPa•s) indicated that S. malaccense may be used as a thickener in hot food., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Background: Leishmaniasis, a neglected disease caused by the parasite Leishmania, is treated with drugs associated with high toxicity and limited efficacy, in addition to constant reports of the emergence of resistant parasites. In this context, snake serums emerge as good candidates since they are natural sources with the potential to yield novel drugs., Objectives: We aimed to show the antileishmanial effects of γCdcPLI, a phospholipase A2 inhibitor from Crotalus durissus collilineatus snake serum, against Leishmania (Leishmania) amazonensis., Methods: Promastigotes forms were exposed to γCdcPLI, and we assessed the parasite viability and cell cycle, as well as invasion and proliferation assays., Findings: Despite the low cytotoxicity effect on macrophages, our data indicate that γCdcPLI has a direct effect on parasites promoting an arrest in the G1 phase and reduction in the G2/M phase at the highest dose tested. Moreover, this PLA2 inhibitor reduced the parasite infectivity when promastigotes were pre-treated. Also, we demonstrated that the γCdcPLI treatment modulated the host cell environment impairing early and late steps of the parasitism., Main Conclusions: γCdcPLI is an interesting tool for the discovery of new essential targets on the parasite, as well as an alternative compound to improve the effectiveness of the leishmaniasis treatment.

The SARS-CoV-2 (COVID-19) virus has caused a devastating global pandemic of respiratory illness. To understand viral pathogenesis, methods are available for studying dissociated cells in blood, nasal samples, bronchoalveolar lavage fluid and similar, but a robust platform for deep tissue characterization of molecular and cellular responses to virus infection in the lungs is still lacking. We developed an innovative spatial multi-omics platform to investigate COVID-19-infected lung tissues. Five tissue-profiling technologies were combined by a novel computational mapping methodology to comprehensively characterize and compare the transcriptome and targeted proteome of virus infected and uninfected tissues. By integrating spatial transcriptomics data (Visium, GeoMx and RNAScope) and proteomics data (CODEX and PhenoImager HT) at different cellular resolutions across lung tissues, we found strong evidence for macrophage infiltration and defined the broader microenvironment surrounding these cells. By comparing infected and uninfected samples, we found an increase in cytokine signalling and interferon responses at different sites in the lung and showed spatial heterogeneity in the expression level of these pathways. These data demonstrate that integrative spatial multi-omics platforms can be broadly applied to gain a deeper understanding of viral effects on cellular environments at the site of infection and to increase our understanding of the impact of SARS-CoV-2 on the lungs., (© 2023 The Authors. Immunology published by John Wiley & Sons Ltd.)

One of the main factors limiting tilapia's production is the occurrence of infections caused by Aeromonas and Streptococcus species. This work intended to evaluate a bivalent vaccine against A. hydrophila and S. agalactiae by intraperitoneal (i.p) administration in Nile tilapia ( Oreochromis niloticus ) in Brazil. The study was carried out in two phases: one in the laboratory, on a small scale, and from the results obtained, the study was expanded to a large scale in a production system in cages. The vaccine proved to be safe and effective in laboratory tests, with a vaccine efficacy (VE) of 93.66%. However, in large-scale tests with 12,000 tilapias, the VE was 59.14%, with a better food conversion ratio (1.54 kg) in the vaccinated group compared to the control group (1.27 kg). These results corroborate the efficiency of this tested vaccine; however, they indicate the need for field tests to attest to real protection.

This work presents a starch extracted from jaboticaba seeds. The extraction yielded 22.65 ± 0.63% of a slightly beige powder (a* 1.92 ± 0.03, b* 10.82 ± 0.17 and L* 92.27 ± 0.24). The starch presented low protein content (1.19% ± 0.11) and phenolic compounds (0.58 ± 0.02 GAE. g) as contaminants. The starch granules showed small, smooth, irregular shapes and sizes between 6.1 and 9.6 µm. The starch presented a high content of amylose (34.50%±0.90) and a predominance of intermediate chain length (B1-chains 51%), followed by A-chains (26%) in the amylopectin. The SEC-MALS-DRI showed the starch had a low molecular weight (5.3·106 g·mol -1 ) and amylose/amylopectin content compatible with a Cc-type starch, confirmed in the X-ray diffractogram. Thermal studies showed a low onset temperature (T0 = 66.4 ± 0.46 °C) and gelatinization enthalpy (ΔH = 9.1 ± 1.19 J g -1 ) but a high-temperature range (ΔT = 14.1 ± 0.52 °C). The jaboticaba starch proved to be a promising material for food and non-food applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

In this study functional properties of a galactose-rich heteropolysaccharide (GH) were accessed. The bands of a galactose-rich polysaccharide were found in FTIR spectra, including those from the fingerprint region. GH was characterized as a dark-red material (L* 25.86 ± 0.75, a* 9.46 ± 1.01, b* 0.65 ± 0.14, Chroma 9.48 ± 1.02) with antioxidant activity of 21.5 ± 0.08, 12.1 ± 0.06 and 0.46 ± 0.04 mmol Trolox Eq/mg GH in FRAP, DPPH and ABTS, respectively. GH presented 44.9% of esterification degree and 10.73 ± 0. 01 mg of GAE/g. The production parameters of GH emulsions (GH concentration, time and ultrasound power) were optimized using a 2 3 Central Composite Rotatable Design (CCRD). Emulsion droplets presented particle size (d µm) varying from 0.823 ± 0.065 to 1.926 ± 0.151, polydispersity index (PDI) from 0.10 ± 0.05 to 0.40 ± 0.01 and zeta potential from -29.25 ± 3.98 to -33.75 ± 1.77. Finally, the high emulsifying activity (EA) (96.67%) and emulsion stability (ES) (97.44%) allow suggesting that GH is a promising polysaccharide for food applications.

In the present study, we aimed to produce CGP/PVA films containing entrapped anti-inflammatory drugs for wound dressing applications. Using a 3 3-1 fractional factorial design, the effect of each component was evaluated on the physicochemical and morphological properties of the produced materials. The best formulation for entrapment of diclofenac sodium and ketoprofen was also determined. The produced films presented high swelling capacity, with some formulations showing o porous structure. CGP/PVA films showed a maximum retention of 75.6% for diclofenac sodium and 32.2% for ketoprofen, and both drugs were released in a controlled manner for up to 7 h. The drug release kinetic was studied, and the data were fitted using a Korsmeyer-Peppas model, which suggested that the release mechanism is controlled by diffusion. These results indicate that CGP/PVA-based matrices have great potential to be used as drug-delivery systems for wound dressing applications, contributing to prolonging the drug's action time and then improving their anti-inflammatory efficacy., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest in the publication., (© King Abdulaziz City for Science and Technology 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Background: Tumor budding (TB) has been investigated in several types of solid tumors. In oral cancer, studies show its association with survival. However, for its implementation in routine histological analyses, results with a high certainty of evidence are needed. Therefore, the aim of this systematic review is to explore the association between tumor budding and overall survival (OS), disease-specific survival (DSS), and disease-free survival (DFS) in oral cancer., Methods: A search was performed in Embase, PubMed, Scopus, Livivo, Web of Science, and Google Scholar. We adopted the following inclusion criteria: studies that evaluate tumor budding in oral cancer, that investigate survival, and presenting cohort design. We excluded reviews and studies without hazard-ratio (HR) data., Results: This systematic review included 22 studies and showed an association between TB and survival. High-grade TB is associated with a worse OS in univariate analysis (HR = 3.11; 95% CI: 2.06-4.69, p<0.01) and multivariate analysis (HR = 2.62; 95% CI: 1.64-4.20, p<0.01); with a poorer DSS in univariate (HR = 2.43; 95% CI: 1.94-3.03, p<0.01) and multivariate analysis (HR = 2.01; 95% CI: 1.43-2.83, p< 0.01); and with a worse DFS in univariate (HR = 1.94; 95% CI: 1.44-2.62, p<0.01) and multivariate analysis (HR = 2.15; 95% CI: 1.31-3.53, p< 0.01). Sensitivity analysis showed that the results are robust, and no significant publication bias was identified in univariate analysis for DFS (Egger's test: p = 0.94). The certainty of the evidence was graded as low or very low., Conclusion: Our findings indicate that TB is an independent prognostic factor of OS, DSS, and DFS in oral cancer. However, further studies are needed to increase the certainty of the evidence.

Recent studies suggest that the association of antigens in microparticles increases the anti- Leishmania vaccine immunogenicity. This study aims to investigate the in situ effect of the adjuvant performance consisting of chitosan-coated poly( D , L -lactic) acid submicrometric particles (SMP) and analyze the inflammatory profile and toxicity. Two formulations were selected, SMP 1 , containing poly( D , L -lactide) (PLA) 1% wt / v and chitosan 1% wt / v ; and SMP 2 , containing PLA 5% wt / v and chitosan 5% wt / v . After a single dose of the unloaded SMP 1 or SMP 2 in mice, the SMPs promoted cell recruitment without tissue damage. In addition, besides the myeloperoxidase (MPO) activity having demonstrated similar results among the analyzed groups, a progressive reduction in the levels of N-acetyl- β- D -glucosaminidase (NAG) until 72 h was observed for SMPs. While IL-6 levels were similar among all the analyzed groups along the kinetics, only the SMPs groups had detectable levels of TNF-α. Additionally, the Leishmania braziliensis antigen was encapsulated in SMPs (SMP 1 Ag and SMP 2 Ag), and mice were vaccinated with three doses. The immunogenicity analysis by flow cytometry demonstrated a reduction in NK (CD3 - CD49 + ) cells in all the SMPs groups, in addition to impairment in the T cells subsets (CD3 + CD4 + ) and CD3 + CD8 + ) and B cells (CD19 + ) of the SMP 2 group. The resulting data demonstrate that the chitosan-coated SMP formulations stimulate the early events of an innate immune response, suggesting their ability to increase the immunogenicity of co-administered Leishmania antigens.

Objective: To recognize changes that occur along the trigeminal pathway in oral cancer in order to establish an effective approach to pain control., Methods: Wistar rats were divided into control and 4-NQO groups for 8, 12, 16, or 20 weeks. 4-NQO suspension was administered on the animals' tongues. Mechanical hyperalgesia, assessment of facial expressions, and an open-field test were performed. After euthanasia, the animals' tongues were removed for macro- and microscopic analysis. c-Fos expression was analyzed in the trigeminal pathway structures., Results: 4-NQO induced time-dependent macroscopic lesions that were compatible with neoplastic tumors. Histopathological analysis confirmed oral squamous cell carcinoma in 50% of the animals on the 20th week. There was a significant nociceptive threshold reduction during the first two weeks, followed by a threshold return to the baseline levels, decreasing again from the 12th week. Facial nociceptive expression scores were observed on the 20th week, while increased grooming and exploratory activity were observed on the 8th week. Trigeminal ganglion showed an increased c-Fos immunoexpression on the 20th week, and in the trigeminal subnucleus caudalis, it occurred on the 16th and 20th. The long-term carcinogenic exposure caused changes in the nociceptive behavior and c-Fos expression in the rats' trigeminal pathway., (© 2022 Wiley Periodicals LLC.)

Objective:  To describe a reference curve for cervical length (CL) in mid-trimester twin gestations using transvaginal ultrasound (TVU) and to investigate whether short CL increases spontaneous preterm birth (sPTB) in asymptomatic twin pregnancies., Methods:  This was a prospective cohort study performed at 17 outpatient antenatal facilities of Brazil with women at 18 0/7 to 22 6/7 weeks of gestation who participated in a randomized clinical trial screening phase (P5 trial) between July 2015 and March 2019. TVU was performed to provide CL measurement in all screened women. Almost all women with CL ≤ 30 mm received vaginal progesterone 200mg/day and they were also randomized to receive cervical pessary or not. We considered data from the CL distribution among asymptomatic twin pregnancies and analyzed CL and its association with PTB generating receiver operating characteristics (ROC) curves and Kaplan-Meier curves., Results:  A total of 253 pregnant women with twins were included in the distribution curve. The mean CL was 33.7 mm and median was 35.5mm. The 10th percentile was 17.8mm. We identified a PTB rate of 73.9% (187/253) with 33.6% of sPTB < 37 (85/253) and 15% (38/253) of sPTB < 34 weeks. The best cutoff point to predict sPTB < 37 was 24.15 mm. However, the ROC curve showed a poor performance (0.64). The Kaplan-Meier survival curves identified that only CL values ≤ 20mm were associated to sPTB < 34 weeks., Conclusion:  A cutoff point of CL ≤ 20 mm can be interesting point to identify short cervix in Brazilian twin pregnancies. However, in Brazilian asymptomatic twin pregnancies, CL does not show a good performance to predict PTB., Competing Interests: The authors have no conflict of interests to declare., (Federação Brasileira de Ginecologia e Obstetrícia. This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/).)

Visceral Leishmaniasis is one of the most important vector-borne zoonoses in the world. In 2013, the first autochthonous canine case of the disease in the state of Paraná, southern region of Brazil, was reported in Foz do Iguaçu, on the triple border between Brazil, Argentina and Paraguay. In 2015, the first human case was related. Once the endemic was confirmed, the Zoonoses Control Center (ZCC), an agency of the Municipal Health Department, started actions to implement the Human Visceral Leishmaniasis (HVL) Surveillance and Control Program (VLSCP), of the Ministry of Health. Between 2015 and 2020, 12,205 dog samples were analyzed for the diagnosis of the disease. A prevalence of 37.94% (4,630 samples) was found: 2016 had the highest prevalence, with 46.25%, and the year with the lowest prevalence was 2020, with 25.98%. Possible risk factors for dogs were analyzed, and the results obtained were: whether the request for the exam was performed by the ZCC was a significant protective factor, with a lower prevalence (37.5%) than dogs coming from private clinics (OR = 0.89, p-value = 0.016). Males were significantly more infected than females, with 41.1% and 35.7% positivity, respectively (OR = 1.24, p < 0.0001). Companion dogs and mixed breed dogs were significantly less affected than the other groups tested (OR = 0.44, p < 0.001; OR = 0.79, p = 0.012, respectively). The dogs' dark coat color was a significant risk factor with respect to the other color categories. Short and medium coat sizes were significantly considered risk factors, with 41.3% and 31.3% positivity. Long-haired dogs had only 22.7% positivity. In univariate analyses, giant, large and medium dogs were significantly more affected than small dogs. Dogs up to four years of age were significantly less affected than those in other age groups. There was a coincidence of human and canine cases in the spatial distribution. However, according to the literature, a higher incidence would be expected in humans, due to the high prevalence found in dogs. Therefore, further studies should be carried out to understand the dynamics of the disease in this region., (Copyright © 2023 Elsevier B.V. All rights reserved.)

In this study, the antioxidant, antimicrobial, mechanical, optical, and barrier attributes of Solanum lycocarpum starch bio-based edible films incorporated with a phenolic extract from jaboticaba peel were investigated. Aiming to determine the effect of the polymers and the phenolic extract on the properties of the films, a three-factor simplex-lattice design was employed, and the formulation optimization was based on the produced films' antioxidant potential. The optimized formulation of the starch-PEJP film showed a reddish-pink color with no cracks or bubbles and 91% antioxidant activity against DPPH radical. The optimized starch-PEJP film showed good transparency properties and a potent UV-blocking action, presenting color variation as a function of the pH values. The optimized film was also considerably resistant and highly flexible, showing a water vapor permeability of 3.28 × 10 -6 g m -1 h -1 Pa -1 . The microbial permeation test and antimicrobial evaluation demonstrated that the optimized starch-PEJP film avoided microbial contamination and was potent in reducing the growth of Escherichia coli , Staphylococcus aureus , and Salmonella spp. In summary, the active starch-PEJP film showed great potential as an environmentally friendly and halochromic material, presenting antioxidant and antimicrobial properties and high UV-protecting activity.

The Web-based Executive Functioning Questionnaire (Webexec) is a brief scale developed to assess executive functions via online format. It has been used in different contexts, but its adaptation to other cultures is still restricted. This study aimed to perform a cross-cultural adaptation of the Webexec for a Brazilian sample considering the psychometric properties of the scale. This study used a sample of 295 Brazilian participants, with a mean age equal to 20.69 ( SD  = 6.030). This is a longitudinal study with reapplication of the scale six weeks after the test phase. Classical and contemporary methods were applied to analyze the psychometric properties of the Webexec. The results showed that the scale presented excellent psychometric properties for the Brazilian version, considering validity evidence based on the content and internal structure of Webexec, as well as reliability and precision. However, it is considered that other relational and experimental studies should be carried out with a larger sample size and for different population groups.

Early diagnosis is essential for the appropriate management of acute kidney injury (AKI). We evaluated the impact of an electronic AKI alert together with a care bundle on the progression and mortality of AKI. This was a single-center prospective study that included AKI patients aged ≥ 18 years, whereas those in palliative care, nephrology, and transplantation departments were excluded. An AKI alert was issued in electronic medical records and a care bundle was suggested. A series of classes were administered to the multidisciplinary teams by nephrologists, and a clinical pharmacist audited prescriptions. Patients were categorized into pre-alert and post-alert groups. The baseline characteristics were comparable between the pre-alert (n = 1613) and post-alert (n = 1561) groups. The 30-day mortality rate was 33.6% in the entire cohort and was lower in the post-alert group (30.5% vs. 36.7%; p < 0.001). Age, pulmonary disease, malignancy, and ICU admission were associated with an increase in 30-day mortality. The electronic AKI alert together with a care bundle and a multidisciplinary education program was associated with a reduction in 30-day mortality in patients with AKI.

Beans reached the research spotlight as a source of bioactive compounds capable of modulating different functions. Recently, we reported antioxidant and oxidonitrergic effect of a low molecular weight peptide fraction (<3 kDa) from hardened bean ( Phaseolus vulgaris ) in vitro and ex vivo , which necessitate further in vivo assessments. This work aimed to evaluate the hypotensive effect and the involved physiological mechanisms of the hardened common bean peptide (Phaseolus vulgaris ) in normotensive (Wistar) and hypertensive (SHR) animals. Bean flour was combined with a solution containing acetonitrile, water and formic acid (25: 24: 1). Protein extract (PV3) was fractioned (3 kDa membrane). We assessed PV3 effects on renal function and hemodynamics of wistar (WT-normotensive) and spontaneously hypertensive rats (SHR) and measured systemic arterial pressure and flow in aortic and renal beds. The potential endothelial and oxidonitrergic involvements were tested in isolated renal artery rings. As results, we found that PV3: I) decreased food consumption in SHR, increased water intake and urinary volume in WT, increased glomerular filtration rate in WT and SHR, caused natriuresis in SHR; II) caused NO- and endothelium-dependent vasorelaxation in renal artery rings; III) reduced arterial pressure and resistance in aortic and renal vascular beds; IV) caused antihypertensive effects in a dose-dependent manner. Current findings support PV3 as a source of bioactive peptides and raise the potential of composing nutraceutical formulations to treat renal and cardiovascular diseases., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

Seminal studies stated that bean proteins are efficient neuronal tracers with affinity for brain tissue. A low molecular weight peptide fraction (<3kDa) from Phaseolus vulgaris (PV3) was previously reported to be antioxidant, non-cytotoxic, and capable of reducing reactive oxygen species and increasing nitric oxide in cells. We evaluated the effects of PV3 (5, 50, 100, 500, and 5000 µg/kg) on behavior and the molecular routes potentially involved. Acute and chronic PV3 treatments were performed before testing Wistar rats: i) in the elevated plus-maze (EPM) to assess the anxiolytic-like effect; ii) in the open field (OF) to evaluate locomotion and exploration; and iii) for depression-like behavior in forced swimming (FS). Catecholaminergic involvement was tested using the tyrosine hydroxylases (TH) enzyme inhibitor, α-methyl-DL-tyrosine (AMPT). Brain areas of chronically treated groups were dissected to assess: i) lipid peroxidation (LPO); ii) carbonylated proteins (CP); iii) superoxide dismutase (SOD) and catalase (CAT) enzymatic activities. Neuronal nitric oxide synthases (nNOS) and argininosuccinate synthase (ASS) protein expression was evaluated by western blotting. Acute treatment with PV3 increased the frequency and time spent in the EPM open arms, suggesting anxiolysis. PV3 increased crossing episodes in the OF. These PV3 effects on anxiety and locomotion were absent in the chronically treated group. Acute and chronic PV3 treatments reduced the immobility time in the FS test, suggesting an antidepressant effect. TH inhibition by AMPT reverted acute PV3 effects. PV3 decreased LPO and CP levels and SOD and CAT activities, whereas nNOS and ASS were reduced in few brain areas. In conclusion, PV3 displayed central antioxidant actions that are concomitant to catecholaminergic-dependent anxiolytic and antidepressant effects.

The COVID-19 pandemic, promoted by the SARS-CoV-2 respiratory virus, has resulted in widespread global morbidity and mortality. The immune response against this pathogen has shown a thin line between protective effects and pathological reactions resulting from the massive release of cytokines and poor viral clearance. The latter is possibly caused by exhaustion, senescence, or both of TCD8+ cells and reduced activity of natural killer (NK) cells. The imbalance between innate and adaptive responses during the early stages of infection caused by SARS-CoV-2 contributes to the ineffective control of viral spread. The present study evaluated the tissue immunoexpression of the tissue biomarkers (Arginase-1, CCR4, CD3, CD4, CD8, CD20, CD57, CD68, CD138, IL-4, INF-α, INF-γ, iNOS, PD-1, Perforin and Sphingosine-1) to understand the cellular immune response triggered in patients who died of COVID-19. We evaluated twenty-four paraffin-embedded lung tissue samples from patients who died of COVID-19 (COVID-19 group) and compared them with ten lung tissue samples from patients who died of H1N1pdm09 (H1N1 group) with the immunohistochemical markers mentioned above. In addition, polymorphisms in the Perforin gene were genotyped through Real-Time PCR. Significantly increased tissue immunoexpression of Arginase, CD4, CD68, CD138, Perforin, Sphingosine-1, and IL-4 markers were observed in the COVID-19 group. A significantly lower immunoexpression of CD8 and CD57 was also found in this group. It is suggested that patients who died from COVID-19 had a poor cellular response concerning viral clearance and adaptive response going through tissue repair.

Patients with COVID-19 may develop symptoms that interfere with food intake. Systemic inflammatory response associated with physical inactivity and/or immobilization during hospital stay can induce weight and muscle loss leading to sarcopenia and worsening the clinical condition of these patients. The present study identifies the frequency and factors associated with sarcopenia prediction in adult and elderly patients hospitalized for COVID-19. It is a cohort-nested cross-sectional study on adult and elderly patients admitted to wards and intensive care units (ICUs) of 8 hospitals in a northeastern Brazilian state. The study was conducted from June 2020 to June 2021. Sociodemographic, economic, lifestyle, and current and past clinical history variables were collected. Sarcopenia prediction was determined by the Strength, Assistance in walking, Rise from a chair, Climb stairs, and Falls (SARC-F) questionnaire compiled in the Remote-Malnutrition APP (R-MAPP). Patients were diagnosed with sarcopenia when the final score ≥ 4 points. The study included 214 patients with a mean age of 61.76 ± 16.91 years, of which 52.3 % were female and 57.5 % elderly. Sarcopenia prevailed in 40.7 % of the sample. Univariate analysis showed greater probability of sarcopenia in elderly individuals, nonpractitioners of physical activities, hypertensive patients, diabetic patients, and those hospitalized in the ICU. In the multivariate model, the type of hospital admission remained associated with sarcopenia prediction, where patients admitted to the ICU were 1.43 (95 % CI: 1.04; 1.97) more likely to have sarcopenia than those undergoing clinical treatment. Sarcopenia prediction was not associated with patient outcome (discharge, transfer, or death) (p = 0.332). The study highlighted an important percentage of sarcopenia prediction in patients with COVID-19, especially those admitted to the ICU. Additional investigations should be carried out to better understand and develop early diagnostic strategies to assist in the management of sarcopenic patients with COVID-19., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Objectives: The present study aimed to identify associations between extremes in body weight status (underweight and excess body weight) before a COVID-19 diagnosis and clinical outcomes in patients infected with SARS coronavirus type 2., Methods: A multicenter cohort study was conducted in eight different states in northeastern Brazil. Demographic, clinical (previous diagnosis of comorbidities), and anthropometric (self-reported weight and height) data about individuals who tested positive for COVID-19 were collected. Outcomes included hospitalization, mechanical ventilation, and death. Multivariable logistic regression models, adjusted based on age, sex and previous comorbidities, were used to assess the effects of extremes in body weight status on clinical outcomes., Results: A total of 1308 individuals were assessed (33.6% were elderly individuals). The univariable analyses showed that only hospitalization was more often observed among underweight (3.2% versus 1.2%) and overweight (68.1% versus 63.3%) individuals. In turn, cardiovascular diseases were more often observed in all clinical outcomes (hospitalization: 19.7% versus 4.8%; mechanical ventilation: 19.9% versus 13.5%; death: 21.8% versus 14.1%). Based on the multivariable analysis, body weight status was not associated with risk of hospitalization (underweight: odds ratio [OR]: 1.10; 95% confidence interval [CI] 95%, 0.50-2.41 and excess body weight: OR: 0.81; 95 CI, 0.57-1.14), mechanical ventilation (underweight: OR: 0.92; 95% CI, 0.52-1.62 and excess weight: OR: 0.90; 95% CI, 0.67-1.19), and death (underweight: OR: 0.61; 95% CI, 0.31-1.20 and excess body weight: OR 0.88; 95% CI, 0.63-1.23)., Conclusions: Being underweight and excess body weight were not independently associated with clinical outcomes in patients with COVID-19 in the herein analyzed cohort. This finding indicates that the association between these variables may be confounded by both age and comorbidities., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Considerable heterogeneity and ontogenetic changes in venom composition have already been observed in different species of snakes within the Viperidae family. Since the venom of young and adult can cause distinct pathological effects and because the antivenom may be less effective in neutralizing envenoming by young snakes compared to adults, it is of paramount importance to understand the ontogenetic variation of snake venom. Thus, the present study aimed to analyze and compare the venom of Bothrops pauloensis snakes, searching for possible influences of ontogeny and sex in their biochemical and biological aspects. The venom of younger individuals was more complex in relation to high molecular mass proteins, with a greater abundance of metalloproteinases, while adults showed a greater abundance of medium and low molecular mass proteins, such as phospholipases A 2 (PLA 2 ), C-type lectins and serine proteases. The antivenom showed better immunorecognition towards the venom of adult snakes than younger ones, in addition to a deficiency in the recognition of medium molecular mass proteins, suggesting the need for an improvement in the antivenom. Younger snakes showed higher coagulant, caseinolytic, and hemorrhagic activity, while adult snakes showed higher L-amino acid oxidase (LAAO) activity and acted faster in lethality. Differences between males and females were observed mainly in the rate of loss of coagulant activity, change in PLA 2 activity and lethality action time. Furthermore, considering only the adult groups, males showed a higher LAAO and thrombin-like activity, while females showed a higher caseinolytic and hyaluronidase activity. With the results obtained in this work, it was possible to conclude that there is an ontogenetic variation in the composition and some activities of the B. pauloensis snake venom, in addition to differences between the venom of males and females, reinforcing that there is an intraspecific variation that may result in different symptoms in their envenoming and, consequently, differences in the response to treatment with the antivenom., (Copyright © 2022. Published by Elsevier Ltd.)

This work compared the presence of phospholipase A 2 inhibitors (PLIs) in the serum of 19 snake species maintained at Instituto Butantan to better understand the mechanisms of venom resistance in snakes and improve the treatment of snakebite. PLI was isolated from blood of 19 snake species by one-step chromatography and identified in all samples, besides its identity was confirmed through the interaction with both phospholipase A 2 and anti-γPLI. These findings highlight the diversity of snake serum PLIs and emphasize the importance of structure-function studies., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Despite coagulotoxicity being a primary weapon for prey capture by Bothrops species (lancehead pit vipers) and coagulopathy being a major lethal clinical effect, a genus-wide comparison has not been undertaken. To fill this knowledge gap, we used thromboelastography to compare 37 venoms, from across the full range of geography, taxonomy, and ecology, for their action upon whole plasma and isolated fibrinogen. Potent procoagulant toxicity was shown to be the main venom effect of most of the species tested. However, the most basal species ( B. pictus ) was strongly anticoagulant; this is consistent with procoagulant toxicity being a novel trait that evolved within Bothrops subsequent to their split from anticoagulant American pit vipers. Intriguingly, two of the arboreal species studied ( B. bilineatus and B. taeniatus ) lacked procoagulant venom, suggesting differential evolutionary selection pressures. Notably, some terrestrial species have secondarily lost the procoagulant venom trait: the Mogi Mirim, Brazil locality of B. alternatus ; San Andres, Mexico locality of B. asper ; B. diporus ; and the São Roque of B. jararaca . Direct action on fibrinogen was extremely variable; this is consistent with previous hypotheses regarding it being evolutionary decoupled due to procoagulant toxicity being the primary prey-capture weapon. However, human patients live long enough for fibrinogen depletion to be clinically significant. The extreme variability may be reflective of antivenom variability, with these results thereby providing a foundation for such future work of clinical relevance. Similarly, the venom diversification trends relative to ecological niche will also be useful for integration with natural history data, to reconstruct the evolutionary pressures shaping the venoms of these fascinating snakes.

Background: Calcium-binding proteins are heterogeneous proteins that act binding this ion in specific domains, performing numerous functions., Objective: In the present review, we aim to gather principal information about S100B protein in the Central Nervous System (CNS), highlighting its particularities, mapping, functionalities, and consequences on CNS dysfunction., Methods: The research was carried out by searching Pubmed, Medline, Science Direct, Lilacs, the Cochrane Library, and Web of Science databases using the following descriptors: S100 protein; Central Nervous System; Nervous Lesions, as well as their corresponding terms in Portuguese and Spanish. The terms were first searched separately, then together., Results: Due to its ability to bind with calcium, S100B is involved in the regulation of several intra- and extracellular physiological processes. As well as being multifunctional, this protein can be considered both a "marker" and "signaling" since it is capable of triggering functions of detection of and protection in situations of injury to the CNS., Conclusions: In-depth studies are necessary to discover the innumerable actions of this protein which are still unknown. It is expected that these can bring varied benefits by elucidating its therapeutic potential in preclinical and clinical situations.

Aiming to understand the impact of hardening on the biological potential of bean protein and peptides, we evaluated the antioxidant and vasorelaxant properties of common beans after and before hardening. It was also evaluated the effect of extrusion and autoclaving in the biological potential of hardened beans. In general, hardening caused a reduction from 13.5 to 39.6% on the antioxidant activity of the peptide-rich fractions. On the other hand, hardening did not strongly interfere with the vascular reactivity in thoracic aorta rings, being observed maximal relation varying from 801% to 84.7%. The thermal treatment caused a general increase in the antioxidant and vasorelaxant potential of these fractions, being observed EC 50 values ranging from 0.22 mg mL -1 to 0.26 mg mL -1 . We can conclude that hardening did not seem to affect definitively the bioactivity of the obtained peptide-rich fractions. Finally, this study allows suggesting practical applications of extrusion as a thermal process in the production of functional food ingredients, and as ready-to-eat products presenting nutraceutical potential. In addition, autoclaving can be used as a pre-treatment of the hardened grains aiming to use them as whole grains with potentialized benefits for human health., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

Polymeric membranes are a viable and sustainable option for the biotechnology industry from an economic and environmental point of view. In this study, we evaluated tissue response and tolerance to the implantation of a polymeric membrane prepared with cashew gum polysaccharide (CGP) associated with polyvinyl alcohol (PVA). The objective was to characterize the biocompatibility of the CGP/PVA membrane in vivo. Following the evaluation criteria of the ISO 10993-6 standard, we demonstrated that the CGP/PVA membrane showed moderate tissue reaction, with a non-irritating ISO pattern, a thinner fibrous capsule, and a smaller amount of collagen compared to the positive control group. At 30 and 60 days, the membrane presented a similar amount of mast cells to that observed in the negative control group. The data demonstrate that the CGP/PVA membrane presents biocompatibility in accordance with the ISO 10993-6 standard.

In this study, a nanocomposite produced with a blend of polyvinyl alcohol and partially hydrolyzed starch from Solanum lycocarpum was used as a matrix to entrap natural bioactive peptides from Phaseolus vulgaris. The nanocomposites were characterized by dynamic light scattering, scanning electron microscopy, and field emission gun scanning electron microscopy. The nanocomposites were then orally administered to Wistar rats, and their absorption was determined using morphometric, histopathological, cytochemistry, transmission electron microscopy, and biochemical analysis. Results showed that despite some aggregates being formed, the nanocomposites efficiently entrapped the natural peptides, with a loading capacity of 303.62 mg (45.7%) and an entrapment efficiency of 85.3% (267.02 µmol). Histochemical and morphological analysis revealed the absence of tissue injury and cellular changes, indicating the absence of deleterious and toxic effects. Transmission electron microscopy showed the internalization of the nanocomposites in the enterocytes, and biochemical analysis indicated that natural peptides were absorbed reaching the bloodstream., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Brain benefits from physical exercise associated with antioxidant supplements such as flaxseed oil. This low cost and simple association may improve hippocampal plasticity, which may work as a preventive and effective therapy in neuroprotection and neuroplasticity processes. This work evaluated the effects of physical exercise with flaxseed oil supplementation (Linum usitatissimum L.) in the hippocampus of Wistar rats. We separated male Wistar rats into four experimental groups: control group (sedentary), a sedentary group with a supplemental diet of flaxseed oil, a group under exercise program with flaxseed oil supplementation, and a group exclusively under exercise program. The swimming exercise consisted of a progressive 28‑day protocol followed by behavioral assessment, brain perfusion, microtomy, immunohistochemistry for glial fibrillary acidic protein (GFAP), cellular morphology, and optical density analysis. We used the ANOVA test with Tukey's post‑test for behavioral analysis. The exercise program with flaxseed oil supplementation was able to alter the GFAP expression in astrocytes in the CA1, CA3 and dentate gyrus regions of the hippocampus and modulate the behavioral aspects of memory and anxiety.

The use of venom in predation exerts a corresponding selection pressure for the evolution of venom resistance. One of the mechanisms related to venom resistance in animals (predators or prey of snakes) is the presence of molecules in the blood that can bind venom toxins, and inhibit their pharmacological effects. One such toxin type are venom phospholipase A 2 s (PLA 2 s), which have diverse effects including anticoagulant, myotoxic, and neurotoxic activities. BoaγPLI isolated from the blood of Boa constrictor has been previously shown to inhibit venom PLA 2 s that induced myotoxic and edematogenic activities. Recently, in addition to its previously described and very potent neurotoxic effect, the venoms of American coral snakes (Micrurus species) have been shown to have anticoagulant activity via PLA 2 toxins. As coral snakes eat other snakes as a major part of their diet, neonate Boas could be susceptible to predation by this sympatric species. Thus, this work aimed to ascertain if BoaγPLI provided a protective effect against the anticoagulant toxicity of venom from the model species Micrurus laticollaris in addition to its ability shown previously against other toxin types. Using a STA R Max coagulation analyser robot to measure the effect upon clotting time, and TEG5000 thromboelastographers to measure the effect upon clot strength, we evaluated the ability of BoaγPLI to inhibit M. laticollaris venom. Our results indicate that BoaγPLI is efficient at inhibiting the M. laticollaris anticoagulant effect, reducing the time of coagulation (restoring them closer to non-venom control values) and increasing the clot strength (restoring them closer to non-venom control values). These findings demonstrate that endogenous PLA 2 inhibitors in the blood of non-venomous snakes are multi-functional and provide broad resistance against a myriad of venom PLA 2 -driven toxic effects including coagulotoxicity, myotoxicity, and neurotoxicity. This novel form of resistance could be evidence of selective pressures caused by predation from venomous snakes and stresses the need for field-based research aimed to expand our understanding of the evolutionary dynamics of such chemical arms race., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Studies that assess the food intake and nutritional status of ostomy patients are scarce in the literature. However, such individuals have symptoms in the postoperative period that determine changes in the intake of calories and nutrients as well as anthropometric variables., Objective: Estimate the energy and nutrient intake of ostomy patients and determine correlations with anthropometric variables., Methods: A cross-sectional study was conducted with ostomy individuals in outpatient follow-up at a reference hospital for postoperative ostomy surgery in the city of Recife, Brazil. Demographic, socioeconomic, clinical, anthropometric, and dietary data were collected through interviews and from patient charts. Statistical analyses were performed with the aid of the Statistical Package for the Social Sciences, version 13.0 for Windows, with the level of significance set at 5% (P≤0.05)., Results: The sample was composed of 100 individuals (54% males) with a mean age of 55.1±15.4 years. Colostomy patients predominated (82%) and had a greater frequency of excess weight compared to ileostomy patients (86.36% versus 13.64%). Median intake was below the Estimated Average Requirements, especially for vitamins A, C, and E. Significant inverse correlations were found between carbohydrate intake and both arm circumference and triceps skinfold (P=0.0302 for each) and a positive correlation was found between protein intake and arm muscle circumference (P=0.0158) in male patients., Conclusion: The present study found significant correlations between macronutrient intake and anthropometric variables indicative of reserves of lean and adipose mass. Moreover, intake was below the recommended values according to sex and age group, especially with regards to vitamins.

Aedes aegypti is a mosquito vector of arboviruses such as dengue, chikungunya, zika and yellow fever that cause important public health diseases. The incidence and gravity of these diseases justifies the search for effective measures to reduce the presence of this vector in the environment. Bioinsecticides are an effective alternative method for insect control, with added ecological benefits such as biodegradability. The current study demonstrates that a chitinolytic enzyme complex produced by the fungus Trichoderma asperellum can disrupt cuticle formation in the L3 larvae phase of A. aegypti, suggesting such biolarvicidal action could be used for mosquito control. T. asperellum was exposed to chitin from different sources. This induction of cell wall degrading enzymes, including chitinase, N-acetylglucosaminidase and β-1,3-glucanase. Groups of 20 L3 larvae of A. aegypti were exposed to varying concentrations of chitinolytic enzymes induced with commercial chitin (CWDE) and larvae cell wall degrading enzymes (L-CWDE). After 72 h of exposure to the CWDE, 100% of larvae were killed. The same percent mortality was observed after 48 h of exposure to L-CWDE at half the CWDE enzyme mixture concentration. Exoskeleton deterioration was further observed by scanning and electron microscopy. Our findings indicate that L-CWDE produced by T. asperellum reflect chitinolytic enzymes with greater specificity for L3 larval biomolecules. This specificity is characterized by the high percentage of mortality compared with CWDE treatments and also by abrupt changes in patterns of the cellular structures visualized by scanning and transmission electron microscopy. These mixtures of chitinolytic enzymes could be candidates, as adjuvant or synergistic molecules, to replace conventional chemical insecticides currently in use., (© 2021 American Institute of Chemical Engineers.)

This investigation focuses on the development and optimization of cashew gum polysaccharide (CGP) nanoparticles grafted with polypropylene glycol (PPG) as carriers for diclofenac sodium. The optimization of parameters affecting nanoparticles formulation was performed using a central composite rotatable design (CCRD). It was demonstrated that the best formulation was achieved when 10 mg of CGP was mixed with 10 μL of PPG and homogenized at 22,000 rpm for 15 min. The physicochemical characterization evidenced that diclofenac was efficiently entrapped, as increases in the thermal stability of the drug were observed. The CGP-PPG@diclofenac nanoparticles showed a globular shape, with smooth surfaces, a hydrodynamic diameter around 275 nm, a polydispersity index (PDI) of 0.342, and a zeta potential of -5.98 mV. The kinetic studies evidenced that diclofenac release followed an anomalous transport mechanism, with a sustained release up to 68 h. These results indicated that CGP-PPG nanoparticles are an effective material for the loading/release of drugs with similar structures to diclofenac sodium.

About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction <3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 µg/mL PV3. Oxidative stress was provoked by 3% H2O2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO- and endothelium-dependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 μg/mL PV3. PV3 at 30 μg/mL increased cell viability, while cytotoxicity was observed only with 250 μg/mL PV3. PV3 at 10 μg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H2O2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release.

The venom color variation of Crotalus durissus terrificus (Cdt) is attributed to the presence of the toxin L-amino acid oxidase (LAAO). During the venom milking routine of Instituto Butantan, we have noticed that most venoms of captive Cdt specimens show a yellowish color, while most venoms of wild specimens are white. Here we describe a comparative analysis of long-term captive (LTC) and recently wild-caught (RWC) Cdt, focusing on LAAO variation. For the identification of LAAO in individual venoms, four different approaches were employed: evaluation of the enzymatic activity, SDS-PAGE, Western blotting, and ELISA. In addition, mass spectrometry analysis was performed using pooled samples. Although some variation among these methodologies was observed, it was possible to notice that the presence of LAAO was significantly higher in the venom of LTC individuals. LAAO was identified in 60-80% LTC specimens and in only 10-12% of RWC specimens. Furthermore, this enzyme accounts for 5.6% of total venom proteins of LTC Cdt pooled venom, while it corresponds to only 0.7% of RWC Cdt pooled venom. These findings strongly suggest that captive maintenance increases the expression of LAAO in Cdt venom., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Maintenance of snakes at Butantan Institute started in the last century, intending to produce a different antivenom serum to reduce death caused by snakebites. Through a successful campaign coordinated by Vital Brazil, farmers sent venomous snakes to Butantan Institute by the railway lines with no cost. From 1908 to 1962, the snakes were kept in an outdoor serpentarium, where venom extraction was performed every 15 days. During this period, the snake average survival was 15 days. In 1963, the snakes were transferred to an adapted building, currently called Laboratory of Herpetology (LH), to be maintained in an intensive system. Although the periodicity of venom extraction remained the same, animal average survival increased to two months. With the severe serum crisis in 1983, the Ministry of Health financed remodeling for the three public antivenom producers, and with this support, the LH could be improved. Air conditioning and exhausting systems were installed in the rooms, besides the settlement of critical hygienic-sanitary managements to increase the welfare of snakes. In the early 1990s, snake survival was ten months. Over the years to the present day, several improvements have been made in the intensive serpentarium, as the establishment of two quarantines, feeding with thawed rodents, an interval of two months between venom extraction routines, and monitoring of snake health through laboratory tests. With these new protocols, average snake survival increased significantly, being eight years for the genus Bothrops , ten years for genus Crotalus and Lachesis, and four years for the genus Micrurus . Aiming the production of venoms of good quality, respect for good management practices is essential for the maintenance of snakes in captivity. New techniques and efficient management must always be sought to improve animal welfare, the quality of the venom produced, and the safety of those working directly with the venomous snakes., Competing Interests: Competing interests: The authors declare that they have no competing interests.

In this study, starches from underground stems of Trimezia juncifolia were evaluated during dry season (DSS), wet season (WSS) and sprouting (SS). Results evidenced that drought stress did not interfere with the yield, amylose content and degree of polymerization (DP) of amylopectin. However, the extraction yield in SS was 58% lower, being observed and increase of 7.5% in the content of amylose, and 13.5% in DP values for SS amylopectin, with a predominance of A-chains. The amount of total sugar, the starch granules size as well as solubility and swelling properties varied as function of the phenological status. Also, starch granules changed from A-type polymorph in DSS and SS to a C A -type in WSS. Nevertheless, it was observed a crystallinity reduction from 56% in DSS to 37.1% in SS. In addition, thermograms evidenced the presence of amylose-lipid complexes, with endothermic transition temperatures being affected by drought stress and sprouting. Finally, results demonstrate that underground stems from T. juncifolia have adaptative strategies involving changes in the morphological and physicochemical properties of the starch granules., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Paracoccidioidomycosis (PCM) is a life-threatening systemic mycosis caused by Paracoccidioides spp. This disease comprises three clinical forms: symptomatic acute and chronic forms (PCM disease) and PCM infection, a latent form without clinical symptoms. PCM disease differs markedly according to severity, clinical manifestations, and host immune response. Fungal virulence factors and adhesion molecules are determinants for entry, latency, immune escape and invasion, and dissemination in the host. Neutrophils and macrophages play a paramount role in first-line defense against the fungus through the recognition of antigens by pattern recognition receptors (PRRs), activating their microbicidal machinery. Furthermore, the clinical outcome of the PCM is strongly associated with the variability of cytokines and immunoglobulins produced by T and B cells. While the mechanisms that mediate susceptibility or resistance to infection are dictated by the immune system, some genetic factors may alter gene expression and its final products and, hence, modulate how the organism responds to infection and injury. This review outlines the main findings relative to this topic, addressing the complexity of the immune response triggered by Paracoccidioides spp. infection from preclinical investigations to studies in humans. Here, we focus on mechanisms of fungal pathogenesis, the patterns of innate and adaptive immunity, and the genetic and molecular basis related to immune response and susceptibility to the development of the PCM and its clinical forms. Immunogenetic features such as HLA system, cytokines/cytokines receptors genes and other immune-related genes, and miRNAs are likewise discussed. Finally, we point out the occurrence of PCM in patients with primary immunodeficiencies and call attention to the research gaps and challenges faced by the PCM field., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The objective of this work was to extract, identify and characterize a galactose-rich heteropolysaccharide (GH) from "jaboticaba" peel. The best conditions to extract the GH according to a 2 3 full-factorial experimental design were 90 °C/30 min/pH 1.0, resulting in a 32.32 % yield using lyophilized sample. The chemical structure analyzed by GC/MS and NMR spectra (HSQC/HSQC-TOCSY) showed that the main chain of GH consists of a (1→4) galactoside branched at carbon 3, containing galactose (67.21 %), glucose (15.78 %), arabinose (9.78 %), rhamnose (2.26 %) and traces of esterified and non-esterified uronic acids. Rheological studies revealed that GH suspensions behave as a Newtonian fluid, with calculated molecular mass of 1.48 × 10 5 Da. The absolute viscosity of 1 % (w/v) aqueous suspension of GH decreased from 25 mPa s to 10 mPa s in NaCl and 7 mPa s in CaCl 2 , indicating the polyelectrolyte character of GH., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Aims: This study aimed to evaluate the toxicity and humoral and cellular immune response of three heterologous vaccines against Leishmania infantum, yet containing synthetic peptides from Leishmania major in the experimental model in hamsters., Methods and Results: Through bioinformatics analyses, two Leishmania major Gp63 peptides were predicted and selected for vaccine formulations. Hamsters were divided into four groups, with each group receiving doses of three vaccine formulations containing HLA-DR1 or HLA-A2 peptides plus Montanide TM or both associated with the adjuvant. The animals received three vaccine doses and were evaluated for toxicity after each dose, in addition to being analysed for the production of antibodies and lymphoproliferation on day 211 after the last vaccine dose. Peptides predicted in association with oily adjuvant induced a humoral response and strong lymphoproliferation to Leishmania infantum antigen-specific stimulation., (© 2020 John Wiley & Sons Ltd.)

Concerning snake venoms, numerous authors worked with different species of Bothrops focusing on the ontogeny of these animals. However, according to PubMed database, no results on studies related to Bothrops jararacussu ontogeny were displayed until now. This fact led us to develop a greater interest in the venom ontogenetic variability of this species, which is little explored so far. Among snakes of the genus Bothrops, B. jararacussu was previously described as the one with highest myotoxic activity. Another peculiarity was also observed in its venom: a low rate of immunogenicity. In addition, its activity is not efficiently neutralized by the specific antibothropic serum. Considering these particularities, we performed an ontogenetic study of B. jararacussu using venom samples from newborns of the same litter (<6 months) and adults (>24 months). Our results identified two distinct profiles in the venom of these animals: young individuals with little PLA 2 K-49 and more proteases; and adults with a lot of the same myotoxic PLA 2, but less proteases. The HPLC and SDS-PAGE profiles corroborated our findings. Adults showed more hemorrhagic activity in vivo than juveniles, while adult males showed less activity when compared to females. In vivo myotoxicity activity was higher in adults than in juveniles. Immune recognition assays showed different results for the distinct venom., (Copyright © 2020 Elsevier Ltd. All rights reserved.)